Division of Biotechnology, Department of Chemistry, Center for Chemistry and Chemical Engineering, Lund University, 22100, Lund, Sweden.
Centre for Analysis and Synthesis, Department of Chemistry, Lund University, Box 124, SE-22100, Lund, Sweden.
Microb Cell Fact. 2023 Jun 29;22(1):120. doi: 10.1186/s12934-023-02130-1.
Biobased 5-(hydroxymethyl)furfural (5-HMF) is an important platform that offers numerous possibilities for upgrading to a range of chemical, material and fuel products. One reaction of special interest is the carboligation of 5-HMF into C compounds, including 5,5'-bis(hydroxymethyl)furoin (DHMF) and its subsequent oxidation to 5,5'-bis(hydroxymethyl)furil (BHMF), due to their potential applications as building blocks for polymers and hydrocarbon fuels.
This study was aimed at evaluating the use of whole cells of Escherichia coli carrying recombinant Pseudomonas fluorescens benzaldehyde lyase as biocatalysts for 5-HMF carboligation, recovery of the C derivatives DHMF and BHMF, and testing the reactivity of the carbonyl groups for hydrazone formation for potential use as cross-linking agents in surface coatings. The effects of different parameters on the reaction were investigated to find the conditions for achieving high product yield and productivity.
The reaction with 5 g/L 5-HMF using 2 g/L recombinant cells in 10% dimethyl carbonate, pH 8.0 at 30 °C resulted in DHMF yield of 81.7% (0.41 mol/mol) at 1 h, and BHMF yield of 96.7% (0.49 mol/mol) at 72 h reaction time. Fed-batch biotransformation generated a maximum DHMF concentration of 53.0 g/L (or 26.5 g DHMF/g cell catalyst) with productivity of 10.6 g/Lh, after five feeds of 20 g/L 5-HMF. Both DHMF and BHMF reacted with adipic acid dihydrazide to form hydrazone that was confirmed by Fourier-transform infrared spectroscopy and H NMR.
The study demonstrates the potential application of recombinant E. coli cells for cost-effective production of commercially relevant products.
生物基 5-(羟甲基)糠醛（5-HMF）是一个重要的平台，为一系列化学、材料和燃料产品的升级提供了众多可能性。其中一个特别有趣的反应是将 5-HMF 卡罗利化为 C 化合物，包括 5,5'-双（羟甲基）呋喃（DHMF）及其随后氧化为 5,5'-双（羟甲基）糠醛（BHMF），因为它们有可能作为聚合物和碳氢燃料的构建块。
本研究旨在评估携带重组荧光假单胞菌苯甲醛裂解酶的大肠杆菌全细胞作为 5-HMF 卡罗利化的生物催化剂的用途，回收 C 衍生物 DHMF 和 BHMF，并测试羰基的反应性，以形成腙，作为表面涂层的潜在交联剂。研究了不同参数对反应的影响，以找到获得高产物收率和生产力的条件。
在 10%碳酸二甲酯中，使用 2 g/L 重组细胞在 30°C 下，5-HMF 浓度为 5 g/L 时，1 小时内 DHMF 产率为 81.7%（0.41 mol/mol），72 小时时 BHMF 产率为 96.7%（0.49 mol/mol）。补料分批生物转化在五次补料 20 g/L 5-HMF 后，最大 DHMF 浓度为 53.0 g/L（或 26.5 g DHMF/g 细胞催化剂），生产率为 10.6 g/L/h。DHMF 和 BHMF 均与己二酸二酰肼反应形成腙，这通过傅里叶变换红外光谱和 H NMR 得到证实。
本研究表明，重组大肠杆菌细胞在具有成本效益的生产商业相关产品方面具有潜在的应用。
